U.S. patent application number 10/597036 was filed with the patent office on 2007-06-28 for separation and cleaning of a suspension comprising magnetic microparticles.
This patent application is currently assigned to QIAGEN INSTRUMENTS AG. Invention is credited to Konstantin Lutze.
Application Number | 20070148785 10/597036 |
Document ID | / |
Family ID | 34744469 |
Filed Date | 2007-06-28 |
United States Patent
Application |
20070148785 |
Kind Code |
A1 |
Lutze; Konstantin |
June 28, 2007 |
Separation and cleaning of a suspension comprising magnetic
microparticles
Abstract
A device (10) for automatically separating the solid and liquid
phase of a suspension (78) and for purifying magnetic
microparticles (76) loaded with organic, in particular molecular
biological or biochemical substances, comprises a process area (12)
with devices, which move in a cyclic manner for transporting the
magnetic microparticles (76) in the x-direction. A first guide (14)
is used for supplying sample containers (P) in the x-direction and
second guides (18) are use for supplying reagent containers (R) in
the y-direction to the process area (12). The second guides (18) in
the y-direction extend at an angle (.alpha.) of 30 to 150.degree.
to the x-direction. A carrier element (24), which can be moved back
and forth in the x-direction, comprises carrier plates (24a, 24b,
24c) which can be lifted and lowered in the z-direction,
individually and together, for magnetic or magnetisable transfer
elements (28) arranged in a matrix shape. The reagent containers
(R) can be positioned according to the grid of the transfer
elements (28) which are preferably configured as rod-shaped
permanent magnets or electromagnets, by introduction into the
process area (12), taking place at an angle (.alpha.) and can be
rejected by ejection in the same direction into a waste collector.
The forward movement of the carrier element (24) in the x-direction
takes place with the use of permanent magnetic rods as transfer
elements (28) with loaded, pulled-up membranes (M) or with the use
of rods-shaped electromagnets with the current switched on. The
backward movement counter to the x-direction takes place with the
use of permanent magnetic rods as transfer elements (28) without
membranes (M) or with the use of rod-shaped electromagnets with the
current switched off.
Inventors: |
Lutze; Konstantin;
(Hombrechtikon, CH) |
Correspondence
Address: |
THADDIUS J. CARVIS
102 NORTH KING STREET
LEESBURG
VA
20176
US
|
Assignee: |
QIAGEN INSTRUMENTS AG
Garstligweg 8
Hombrechtikon
CH
CH-8634
|
Family ID: |
34744469 |
Appl. No.: |
10/597036 |
Filed: |
January 4, 2005 |
PCT Filed: |
January 4, 2005 |
PCT NO: |
PCT/CH05/00001 |
371 Date: |
July 7, 2006 |
Current U.S.
Class: |
436/526 |
Current CPC
Class: |
G01N 2035/0415 20130101;
G01N 35/0098 20130101; B03C 1/286 20130101; G01N 35/026 20130101;
Y10S 436/806 20130101; Y10T 436/25375 20150115; G01N 33/54326
20130101 |
Class at
Publication: |
436/526 |
International
Class: |
G01N 33/553 20060101
G01N033/553 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 8, 2004 |
CH |
00024/04 |
Claims
1. Device (10) for the automatic separation of the solid and liquid
phase of a suspension (78) and for purifying magnetic
microparticles (76) loaded with organic, in particluar molecular
biological or biochemical substances, which device (10) comprises a
process area (12) with mechanisms which move in a cyclic manner for
transporting the magnetic microparticles (76) in the x-direction,
characterised in that a first guide 914) is arranged for supplying
sample containers (P) in the x-direction and second guides (18) are
arranged for supplying reagent containers (R) in the y-direction to
the process area (12), wherein the second guides (18) extend in the
y-direction at an angle (.alpha.) of 30 to 150.degree. to the
x-direction, a carrier element (24), which can be moved back and
forth in the x-direction, comprises carrier plates (24a, 24b, 24c)
which can be lifted and lowered in the z-direction, individually
and together, for magnetic or magnetisable transfer elements (28)
which are arranged in a matrix shape, the reagent containers (R)
can be positioned according to the grid of the transfer elements
(28) by introduction, taking place at an angle (.alpha.), into the
process area (12) and can be rejected by ejection in the same
direction into a waste collector.
2. Device (10) according to claim 1, characterised in that the
transfer elements (28) are configured as preferably rod-shaped
permanent magnets or electromagnets.
3. Device according to claim 1 or 2, characterised in that the
lowermost part of the transfer elements (28) dipping into the
sample (P) and reagent containers (R) is covered by a membrane (M)
which can be lifted and lowered, can be deposited and taken off by
means of a relative movement with respect to the transfer elements
(28), and is preferably tubular or beaker-shaped.
4. Device (10) according to any one of claims 1 to 3, characterised
in that the angle (.alpha.) between x- and y-direction is
90.degree..
5. Device according to any of the claims 1 to 4, characterised in
that the relative movement of a transfer element (28) to the
corresponding membrane (M), in the longitudinal direction (z)
thereof, takes place by means of different lifting or lowering of
corresponding carrier plates (24b, 24c) and of the membranes
(M).
6. Device (10) according to any of the claims 1 to 5, characterised
in that a third guide is arranged for continuously supplying the
membranes (M) at an angle (.beta.) of 60 to 120.degree. with
respect to the x-direction.
7. Device (10) according to any of the claims 1 to 6, characterised
in that a carrier block (46) with channels (48) for the reagent
containers (R), which channels extend perpendicularly to the
x-direction, is arranged in the process area (12) and each have two
horizontal grooves (66) in the side walls (47) which are opposing
at the same level and open on the end face.
8. Device (10) according to claim 6, characterised in that beams
(72) which can be displaced horizontally back and forth, comprising
permanent magnets (74) arranged in the region of the lowerable
transfer elements (28), are arranged in recesses (50) extending
parallel to the channels (48), to resuspend and mix the
microparticles (76).
9. Sample and reagent container (P, R) and membranes (M) for use in
a device (10) according to any of the claims 1 to 8, characterised
in that they are configured as substantially strip-shaped,
stackable cassettes with a plurality of beaker-shaped cavities (22,
36) corresponding to the grid of the transfer elements (28) in the
carrier element (24).
10. Sample and reagent container (P, R) according to claim 9,
characterised in that the preferably six to ten cavities (22) are
preferably flat or oval in cross-section, their cross-sectional
internal diameters preferably being only a little larger that the
corresponding dimensions of the transfer elements (28) or the
pulled-on membranes (M).
11. Method for automatically separating the solid and liquid phase
of a suspension and purifying the solid phase comprising a device
(10), sample containers (P) and reagent containers (R) according to
any one of claims 1 to 10, characterised in that the forward
movement of the carrier element (24) in the x-direction takes the
place with the use of permanent magnetic rods as transfer elements
(28) with loaded, pulled-up membranes (M) or with the use of
rod-shaped electromagnets with the current switched on, and the
backward movement counter to the x-direction takes place with the
use of permanent magnetic rods as transfer elements (28) without
membranes (M) or with the use of rod-shaped electromagnets with the
current switched off.
12. Method according to claim 11, characterised in that the filled
sample containers (P) are firstly guided intermittently or
continuously on the longitudinal side in the x-direction and the
reagent containers (R) with different or at most partially the same
fillings are guided continuously in the y-direction ant the end
face to the process area (12), on each initiation of a new
operating cycle, one membrane (M) in each case is put over the
rearmost transfer elements (28) in the x-direction, the latter are
lowered into the sample container (P) disposed at the process area
(12) and, after attachment of the magnetic microparticles (76) to
the membrane (M), the transfer elements (28) with the membrane (M)
are raised from the suspension liquids, the carrier element (24) is
displace forward in the x-direction by a grid unit, corresponding
to the spacing (a) between two reagent containers (R), the
particle-free sample container (P) is ejected into a waste
container, the filled reagent containers (R) are simultaneously
introduced into the process area (12), the carrier element (24)
with the transfer elements (28) are pulled up of the membranes (M),
the attached magnetic microparticles (76) are resuspended, the
suspension (78) mixed, the transfer element returned by the spacing
(a) counter to the x-direction, while the membranes (M) remain in
their position.
13. Method according to claim 11 or 12, characterised in that upon
each movement of the carrier element (24) in the x-direction, the
membranes (M) are entrained by one grid unit and are ejected at the
end of the process area (12) into a waste collector.
14. Method according to any one of claims 11 to 13, characterised
in that the last reagent container (R) in the x-direction, which is
ejected from the process area (12), is supplied for a further
use.
15. Method according to any one of claims 11 to 14, characterised
in that a working cycle lasts 2 to 4 min.
16. Method according to any one of claims 11 to 15, characterised
in that the reagent containers (R) are used with different
reagents, but with the same reagent in all the cavities (22) of the
same reagent container (R).
Description
[0001] The invention relates to a device for the automatic
separation of the solid and liquid phase of a suspension and for
purifying magnetic microparticles loaded with organic, in
particular molecular biological or biochemical substances, which
device comprises a process area with mechanisms which move in a
cyclic manner for transporting the magnetic microparticles in the
x-direction. The invention also relates to sample and reagent
containers for use in the device and a method for automatic
separation and purification in the device.
[0002] The investigation and/or analysis of organic, in particular
molecular biological substances by determining their chemical or
physical properties with specifically developed methods is
continuously increasing in importance. The use of an inert carrier
in the form of microparticles is advantageous for the chemical
analysis of molecular biological substances, for example blood or
urine. If these microparticles consist of a magnetic or
magnetisable material, contain a material such as this or are
covered with such a material, the solid phase can be separated off
from a suspension with a magnetic field and isolated by subsequent
purifying or washing processes with a very high degree of
cleanliness. Non-magnetic microparticles are sedimented, aspirated
off or decanted, which makes comparatively complicated,
long-lasting and/or frequent washing processes with at least one
buffer solution necessary.
[0003] Loaded magnetic microparticles are separated off according
to known methods, in particular, in that they are deposited by
permanent magnets on the wall of a reaction vessel with the
formation of a cluster and are fixed there during pipetting or
decanting of the suspension liquid. During the removal of the
suspension liquid, the magnetic field has to be maintained, which
generally results in complicated methods and devices.
[0004] A method and a device for separating and washing magnetic
solid particles which are arranged finely distributed in liquid
samples is described in DE 3926462 A1.
[0005] The solid particles are loaded with organic substances as
the precursor for a photometric or radiometric evaluation of
patient samples in the case of immunoluminometric and
immunoradiometric tests. The liquid samples are exposed in test
tubes to a magnetic field generated by permanent magnets with the
magnetic solid particles being attached to the inner wall of the
test tubes. After a predetermined time, the residual liquid is
aspirated off while maintaining the magnetic field. The separation
and washing process is carried out fully automatically in a
continuous process and a plurality of permanent magnets are
arranged on a conveying section for the test tubes.
[0006] The separation method for depositing magnetic microparticles
is improved according to EP 0806665 B1, in that according to one
variant, the microparticles which are deposited on the vessel wall
under the effect of a permanent magnet can be resuspended, after
the rinsing water has been aspirated off, with fresh water or a
fresh reagent, and this substantially increases the purification
effect. After a certain dwell time, the microparticles are again
attached under the magnetic effect of the wall of the reaction
vessel and the rinsing water is aspirated off again. This process
can be repeated many times.
[0007] U.S. Pat. No. 6,207,463 B1 disclosed a separation of
magnetic microparticles from a suspension, in that a rod-shaped
permanent magnet, which is covered by a protective layer except for
the point, is dipped into the suspension with magnetic
microparticles. The magnetic microparticles attach to the point of
the transfer element and can be removed from the suspension
solution when the rod is lifted, the cluster adheres to the rod and
can be dipped into a new liquid phase. Thus pipetting or decanting
can be omitted.
[0008] The inventor has set himself the object of providing a
device and a method which allow a fully automatic process sequence
and can be applied easily, rapidly, reliably and economically.
[0009] With reference to the device, the object is achieved
according to the invention in that a first guide is arranged for
supplying sample containers in the x-direction and second guide are
arranged for supplying reagent containers in the y-direction to the
process area, wherein the second guides extend in the y-direction
at an angle .alpha. of 30 to 150.degree. to the x-direction, a
carrier element, which can be moved back and forth in the
x-direction, comprises carrier plates which can be lifted and
lowered in the z-direction, individually and together, for magnetic
or magnetisable transfer elements which are arranged in a matrix
shape, the reagent containers can be positioned according to the
grid of the transfer elements by introduction, taking place at an
angle .alpha., into the process area and can be rejected by
ejection in the same direction into a waste collector.
[0010] Special and developing embodiments of the invention are the
subject of dependent claims.
[0011] Transfer elements are preferably configured as permanent
magnetic rods or as rod-shaped electromagnets.
[0012] The lowermost part of the transfer elements, which dip into
the sample and reagent containers, is expediently covered with a
membrane which can be lifted and lowered, can be deposited and
taken off by a relative movement with respect to the transfer
elements and is preferably tubular or beaker-shaped. the membrane
can be omitted in the case of electromagnetically operating
transfer elements.
[0013] According to the prior art, cyclic batches constantly run
one-dimensionally in the x-direction. According to the invention,
the cyclic batches run two-dimensionally, the sample containers in
the x-direction and the reagent containers, in contrast, in the
y-direction. As is conventional in the case of space coordinates,
the two directions preferably have an angle .alpha. of 90.degree.,
and they also run at right angles with respect to the third,
vertical, space coordinate z. The first guide for supplying the
sample containers in the x-direction is given; it runs in the same
direction as the back and forth movement of the carrier element. In
special embodiments on the other hand, the y-direction for the two
guides may vary in a relatively large angle range. The second
guides expediently extend in parallel but they may also spread out
and/or, rising, from a side. Closed reagent containers, however, at
the latest directly before the process area, have a horizontal
position, so they can be loaded with a reagent or a possible
closure can be torn off or penetrated without risk of the reagent
leaking.
[0014] The relative movement of a transfer element in the
longitudinal direction thereof compared to the membrane preferably
takes place by different lifting and lowering of the relevant
carrier plates or guides with means which are known per se.
Obviously, the relative movement could partially also take place by
means of lifting and lowering the carrier block located below the
reagent containers, but this seems less advantageous.
[0015] The transportation of the magnetic microparticles in the
x-direction preferably takes place, as mentioned, on tubular or
breaker-shaped cavities of the membrane in the lowermost region of
the transfer elements when the forward movement of the carrier
plates takes place in the x-direction and the pulled-up membrane
can be lowered into the immediately following reagent container.
These membranes are expendable materials; they are guided to the
inlet side of the process area, positioned during a lowering
movement, placed on the rearmost transfer elements in the
x-direction and entrained. It must be possible to position the
membranes in the reagent containers, about halfway up the maximum
lifting height of the transfer elements, with and without an
introduced transfer element. For the continuous supply of membranes
to the inlet side of the process area, a third guide is provided
which, with respect to the x-direction, has an angle of preferably
60 to 120.degree.. The membranes are designed such that they can be
introduced into the sample and reagent containers; in other words
they are the same, in particular, with respect to number and form
of the cavities. The membranes are also designed, like the sample
and reagent containers, preferably as injection mouldings or
deep-drawn parts made of plastics material.
[0016] The containers and the membranes are substantially
strip-shaped, stackable cassettes with a plurality of beaker-shaped
cavities corresponding to the grid of the transfer elements in the
carrier element.
[0017] With regard to the method for automatically separating the
solid and the liquid phase of a suspension and for purifying the
solid phase, the object of the invention is achieved in that the
forward movement of the carrier element in the x-direction takes
place with the use of permanent magnetic rods as transfer elements
with loaded, pulled up membranes or with the use of rod-shaped
electromagnets with the current switched on, and the backward
movement counter to the x-direction takes place with the use of
permanent magnetic rods as transfer elements without membranes or
with the use of rod-shaped electromagnets with the current switched
off. Special and developing embodiments of the method are the
subject of dependent claims.
[0018] Firstly, the filled sample containers are preferably guided
intermittently or continuously on the longitudinal side in the
x-direction and the reagent containers with different or at most
partially the same fillings are guided continuously in the
y-direction at the end face to the process area. With each
initiation of a new operating cycle, one membrane in each case is
put over the rearmost transfer elements in the x-direction,
configured as permanent magnetic rods, the latter are lowered into
the sample container disposed at the process area and, after
attachment of the magnetic microparticles to the membrane, the
transfer elements with the membrane are raised from the suspension
liquids. The carrier element is displace forward in the x-direction
by a grid unit, corresponding to the spacing between two reagents
containers, the particle-free sample container is ejected into a
waste container. The filled reagent containers are simultaneously
introduced into the process area, the carrier element with the
transfer elements is lowered into the reagent container, the
transfer elements are pulled out of the membranes, the attached
magnetic microparticles are resuspended and the suspension mixed.
The transfer elements are returned by the spacing a counter to the
x-direction, while the membranes remain in their position.
[0019] On each movement of the carrier plates in the x-direction,
the membranes are entrained by one grid unit, the spacing a, and at
the end of the process area, ejected into a waste container. The
last reagent container in the x-direction, ejected from the process
area, is supplied for any further use which is known per se, for
example chemical analysis.
In the case of transfer elements which are configured as rod-shaped
electromagnets, without a membrane, the current is switched on for
loading with microparticles and switched off for suspension.
[0020] One working cycle preferably lasts 2 to 4 min. The duration
of one working cycle is as low as possible for economic reasons,
currently about 2 min can be achieved.
[0021] With a full working load, 6 to 10 reagent containers are
simultaneously pushed into the process area. All the reagent
containers preferably have different reagents, with pure water or
an organic solution also being called a reagent. Obviously,
however, sequences with individual ore repeating reagents my be put
together. Within the same reagent container, the cavities always
have the same reagent. If not all the channels (48 in FIG. 17) are
occupied by reagent containers, the frontmost channels in the
x-direction remain empty. The rejection of the membranes and the
further use of the frontmost reagent container takes place as if it
were in the frontmost channel.
[0022] The microparticles used, as the name states, have dimensions
from one to a plurality of micrometres and they may also have
fractions of a micrometre and should then correctly be called
nanoparticles. For the sake of simplicity, however, the term
microparticles will be used for all particle sizes. the cavities of
the sample and reagent containers generally have a volume of 1 to 3
ml.
[0023] The advantages of the invention can be summarised as
follows. [0024] The device according to the invention can be
operated fully automatically. [0025] the method with the working
cycles allows all the cavities to be in action during the entire
process. [0026] Numerous modules, generally six to ten, can operate
simultaneously, which means maximum working productivity. [0027]
The matrix arrangement allows as optimally dense arrangement,
resulting in a further increase in productivity. [0028] The method
allows continuous processing of samples.
[0029] The invention will be described in more detail with aid of
embodiments shown in the drawings, which are also subject of
dependent claims. In the drawings, schematically:
[0030] FIG. 1 shows a perspective view of the device,
[0031] FIG. 2 shows a vertical section in the x-direction through
the process area,
[0032] FIG. 3 shows a layout of the device at the beginning of the
process,
[0033] FIG. 4 shows a vertical section in the x-direction through
the process area with the positioned sample container,
[0034] FIG. 5 shows a subsequent process step according to FIG. 4,
with the permanent magnetic rods dipped in the sample container,
with membranes,
[0035] FIG. 6 shows a next process step according to FIG. 5 with a
carrier element displace in the x-direction,
[0036] FIG. 7 shows a layout of the device after filling of the
first reagent container,
[0037] FIG. 8 shows a further process step according to FIG. 6 with
the permanent, magnetic rods pulled up and the membrane in the
first reagent container,
[0038] FIG. 9 shows a further variant with a carrier element
returned in the counter direction to the x-direction,
[0039] FIG. 10 shows a further method step according to FIG. 9 with
the second membrane in place,
[0040] FIG. 11 shows a further method step according to FIG. 10
with lowered permanent magnetic rods,
[0041] FIG. 12 shows a further layout with an ejected first reagent
container,
[0042] FIG. 13 shows a further layout of the device with the
carrier element moved forward in the x-direction,
[0043] FIG. 14 shows a further method step according to FIG. 13
with the carrier element displaced in the x-direction,
[0044] FIG. 15 shows a further layout after a plurality of method
steps with the first reagent container in the end position,
[0045] FIG. 16 shows a last layout with an ejected reagent
container and membrane,
[0046] FIG. 17 shows a partially cut away perspective view of a
carrier block,
[0047] FIG. 18 shows a horizontal section through a magnetic
mixer,
[0048] FIG. 19 shows the magnetic mixer according to FIG. 18 in the
other position, and
[0049] FIG. 20 shows the lowermost region of cavity of a sample or
reagent container.
[0050] A perspective view of a device 10 according to the invention
with the preferably right-angled space coordinates x, y and z
substantially comprises a central process area 12, where the
separation and purifying processes take place, first guides 14,
indicated only by dashed lines, for sample containers P and second
guides 18 for reagent containers R. The first guides 14 in the
x-direction and the second guides 18 in the y-direction have an
angle .alpha. of 90.degree., in other words extend at right angles.
Both the samples P and the reagent containers R are substantially
strip-shaped and have tubular or breaker-shaped cavities 22, which
are produced by injection moulding from plastics material, the
sample containers P and reagent containers R being identical. A
peripheral flange 16 does not only stabilise the cavities 22, it is
also used for guidance and holding.
[0051] The process area 12 is limited in the horizontal extent,
largely by a carrier element 24, which consists of three carrier
plates 24a, 24b and 24c. The entire carrier element 24 is lifted
and lowered in the z-direction with the lowermost carrier plate
24a, and the drive and the control for this are implemented with
means which are known per se, like the drive for supplying the
sample containers P and the reagent containers R. The carrier
plates 24b and 24c can be lifted and lowered together, but also
individually, resulting in a relative displacement. In this case,
the transfer elements 28, which are configured as permanent
magnetic rods, are pushed into the membranes M or pulled out
therefrom.
[0052] The carrier element 24 has a predetermined grid of holes 30,
which are penetrated by permanent magnetic rods 28. These have
peripheral collars, which rest on the carrier plate 24c. The
membranes M are basically configured like the sample containers P
and reagent containers R, but the cavities 32 are generally
cylindrical. A prepared membrane M is supplied from the front left
with a third guide, which is not shown for the sake of clarity. The
membranes M are received at the entry to the process area 12 and
conveyed during each working cycle by the grid distance a in the
x-direction. This takes place by means of displacement on a
horizontal pair of rails 36.
[0053] During each working cycle one filled sample container P, on
the longitudinal side, reaches the process area 12. Magnetic
microparticles (76 in FIG. 20) attach to the membranes M or its
cavities 32 at the frontmost sample container P in the x-direction,
when the permanent magnetic rods 28 are introduced. These
microparticles are conveyed stepwise from reagent container R to
reagent container R by way of the membranes M, with it being
possible to resuspend the attached microparticles in each working
cycle. The sample containers P are ejected in the y-direction and
collected in a waste container, not shown.
[0054] The reagent containers R which are supplied in the
y-direction have also been filled in-line. If preassembled, filled
reagent containers R are used, these are torn open or penetrated in
the lid region directly before the process area 12, so a membrane M
or the permanent magnetic rods 78 can be supplied. The mechanisms
for filling or opening are arranged in or on a housing 38 which can
also be lifted or lowered in the z-direction. In the working cycle,
in the present case, six reagent containers R are introduced into
the process area 12 and the used containers are ejected in the
y-direction into a waste collector. Neither sample containers P nor
reagent containers R are guided in the x-direction through the
process area 12.
[0055] In each working cycle of about three minutes in duration, 48
samples are simultaneously separated or purified or washed. Per
working cycle, eight samples leave the process 12, in other words
about 160 samples in an hour.
[0056] In FIG. 2, the process area 12 is shown at the beginning of
a working cycle. The carrier plates 24a and 24b are lifted to such
an extent that the membranes M are located above the level of the
suspension 40 in the reagent containers R. The uppermost carrier
plate 24c is lifted to such an extent that the permanent magnetic
rods 28 are practically pulled out of the membranes M. The
frontmost sample container P in the x-direction, the advance
direction, is still outside the process area 12.
[0057] A holder 34 with a membrane M is moved to the process area
12, and the membrane M can be put over the permanent magnetic rods
28 in the z-direction. This is the first process step of a working
cycle. The path of the membranes M through the entire process 12 in
the x-direction is indicated by arrows 44.
[0058] Channels 48 for the reagent containers R which are inserted
from the rear, positioned in the working position according to the
grid of the permanent magnetic rods 28, and ejected to the front,
which channels are open on either side, extend perpendicularly to
the x-direction in the carrier block 46. Configured in an
alternating manner with respect to channels 48, which are open
laterally and upwardly are recesses 50, in which beams 52 which can
be pushed back and forth, i.e. perpendicularly to the drawing
plane, in the y-direction, are arranged, with permanent magnets,
shown later in detail, for mixing the suspension 40.
[0059] A layout according to FIG. 3 shows--viewed from the top--the
process area 12, in which neither sample containers P, nor reagent
containers R, nor membranes M are introduced. A sample container P1
placed according to the grid in x-direction at a spacing a is
covered by a membrane M1, which is put over the permanent magnetic
rods 28 from below. A further section 53 contains the sample
containers P2 to P7, which are fed in by filling station 54.
[0060] In the y-direction, six reagent containers R1.1 to R6.1 have
been pushed at the end face to the process area 12. In this
position, the reagent containers R are opened or filled with
reagents. The reagent containers R of a buffer section 58 are
designated by R2.1 to R6.2.
[0061] FIG. 4 shows the situation according to FIG. 3 in vertical
section in the x-direction. The rearmost permanent magnetic rods 28
in the x-direction have the membrane M1 put over from below. In the
x-direction, the frontmost sample container P1 is lined up at the
process area 12. The latter is located precisely below the membrane
M1. The holder 34 of the membrane M1 is ejected and disposed of
after the membrane M1 has been lifted, and this is indicated by an
arrow.
[0062] In FIG. 5 the par of rails 36 and the carrier element 24 are
completely lowered in the direction z and the membranes M1 are
dipped with the inserted permanent magnetic rod 28 into the
suspension of the sample container P1 placed on the longitudinal
side at the process area 12. Under the action of the permanent
magnetic rods 28, the magnetic microparticles of the suspension
settle on the membrane M1 and remain suspended on the membrane 26
when the pair of rails 36 and the carrier element 24 are
raised.
[0063] FIG. 6 shows the pair of rails 36 and carrier element 24
which have been raised to the same degree. In this position, the
carrier element 24 has been displaced in the x-direction by the
grid spacing a. The rearmost permanent magnetic rods 28 in the
x-direction with the membrane M1 placed over are now precisely
above the reagent container R.1.1 which has been introduced in the
meantime. The membrane M1 has been pushed into this position
according to FIG. 6 in the pair of rails 36. The reagent container
1.2, according to FIG. 7, has slipped from the buffer section 58
into the buffer section 52 where it is filled with reagents or the
seal is pushed in. The reagent container R1.3 has been moved up
onto the buffer section 58.
[0064] Compared to the layout according to FIG. 3, the sample
container P1 has been pushed away in the y-direction and falls into
a waste collector, not shown. This is indicated by an arrow 60.
[0065] It is shown in FIG. 8 that the carrier element 24 with the
carrier plates 24a, 24b and 24c are lowered to such an extent that
the first membrane M1 has been dipped into the reagent container
R1.1. Under the magnetic effect, the microparticles collect on the
surface of the membrane M1. After a reaction time of about 1
minute, the carrier plate 24c with the permanent magnetic rods 28
is pulled out of the membrane M1. A magnetic mixing mechanism 62 is
now started up. The magnetic microparticles are released by the
mixing mechanism 62 from the membrane M1 and are resuspended.
[0066] In the next working step according to FIG. 9, the complete
carrier element 24 is raised until the permanent magnetic rods 28
are completely removed from the membrane M1 which was also raised
with the pair of rails 36. A second membrane M2 is then provided.
The carrier element 24 can now be returned counter to the
x-direction by one grid unit a and the rearmost permanent magnetic
rods 28 in the x-direction are now precisely above the provided
second membrane M2.
[0067] In the subsequent method step according to FIG. 10 the
carrier element 24, consisting of three carrier plates 24a, 24b and
24c, are moved down in the z-direction, until the second to
rearmost permanent magnetic rods 28 in the z-direction have been
dipped into the first membranes M1 in the working position. At the
same time, the second membranes M2 are put over the permanent
magnetic rods 28.
[0068] In FIG. 11, three process steps are combined. Finally, the
holder 34 for the second membrane M2 is removed, which is
characterised by an arrow. then, the second to frontmost sample
container P2 in the x-direction is guided to the process area 12,
and finally the carrier 24 is lowered to such an extent that the
first membrane M1 is dipped into the first sample container P1.1
and the second membrane M2 is dipped into the second to frontmost
sample container P2 and the corresponding permanent magnetic rods
28 have been lowered into the membrane M2. The microparticles are
now attached to the two first membranes M1, M2. When the permanent
magnetic rods 28 are pulled up, they can be removed from the liquid
phase of the suspension. The microparticle-free reagent container
R1.1 can now be pushed out of the process area 12 in the
y-direction and this is shown in the layout of FIG. 12. Only the
first membrane M1 remains in the process area 12.
[0069] According to the following layout shown in FIG. 13, the
second sample container P2 is also ejected in the y-direction and
fed to the waste collector. The process containers R1.2 and R2.1
are pushed up into the process area 12 and positioned below the
membranes M1 and M2.
[0070] According to FIG. 14, the carrier element 24 with the
permanent magnetic rods 28 and the membranes M1 is pushed forward
by a grid unit a in the x-direction and then lowered as a whole in
the z-direction until the membranes M1 and M2 are positioned in the
two first reagent containers R1.2 and R2.1. The magnetic
microparticles are now collected on the outer wall of the membranes
M1 and M2 and are attached. In the new working cycle started with
FIG. 9, the procedure is now as in the previous working cycle:
lifting the permanent magnetic rods 28, mixing the releasing
magnetic microparticles etc. Each new working cycle is started with
placing of a membrane M and the removal of the samples from the
frontmost sample container P lined up in the x-direction, on the
longitudinal side at the process area 12.
[0071] In the layout according to FIG. 15, the membrane M1 has
reached the last working position in the x-direction in the process
area 12. The treatment of the samples has been competed; all the
provided operations have then been carried out. Only in this
configuration is the device fully operable.
[0072] In the layout according to FIG. 16, it is shown that the
first membrane M1, which has run through all the working cycles, is
fed to the waste collector after ejection from the process area 12.
The reagent container 6.1 with the result of the six working
cycles, on the other hand, is supplied for use, which is indicated
by the arrow 64.
[0073] FIG. 17 shows a carrier block 46, which has been broken open
for the sake of clarity, of the process area 12. There are six
continuous channels 48 which are open at the top, for the message
of the reagent containers R. When passing through, the reagent
containers R slide with their peripheral flange 16 in opposing
grooves 66 in the side walls 47 of the channels 48.
[0074] Provided between the channels 48 of the carrier block 46 are
further recesses 68, which alternate with the channels 48 and are
closed on the end face 70 of the carrier block 46. The recesses 68
are used for receiving beams 72, which can be pushed back and forth
in the y-direction, with integrated permanent magnets 74. The
permanent magnets 74 are arranged in the region of the cavities 22
of the reagent containers R and are used for mixing the resuspended
magnetic microparticles 76.
[0075] As can be seen from FIGS. 18 and 19, the beans 72a, 72b
which can be moved back and forth are arranged on either side of
cavities 22 of a reagent container R. The permanent magnets 74 are
arranged in the double spacing of the cavities 22. When switched
on, there is a relative movement between the permanent magnets 74
and the cavities 22. The microparticles 76 which are attached on
the side, change side, so an effective mixing effect is produced.
The effect can be improved in that the cavities are configured so
as to be elliptical, oval or rectangular with round short
sides.
[0076] FIG. 20 shows a greatly enlarged view of the lower region of
a cavity 22 with a suspension 78. The lowermost region of a
permanent magnetic rod 28 is covered with a beaker-shaped membrane
M. The permanent magnets 28 have the effect that the microparticles
76 collect on the membrane M or the cavities 32. If the membrane M
is removed together with the permanent magnetic rods 28, the
microparticles 76 are lifted from the practically particle-free
suspension 78. If, on the other hand, the permanent magnetic rod 28
is removed and the membrane M left, the microparticles 76 are
released again from the membrane M. By mixing, for example as shown
in FIGS. 18 and 19, the release process can be accelerated and the
mixing effect improved.
* * * * *